Project Summary Treatment of immunological disorders with emerging treatments such as immunotherapy represent a growing medical paradigm. To create potent and targeted therapies, defining the underlying immunological mechanisms of action and is critical. Because of immunological complexity, measurements across multiple immune populations are essential as both diseases and treatments are often driven by several subpopulations of cells. However, due to limitations with current cell isolation and analysis tools, precisely mapping these multi-population interactions is difficult and cost prohibitive to deploy at a scale conducive for high throughput target discovery or streamlined clinical monitoring. Most high precision cell isolation and analysis tools, such as FACS or microfluidic platforms, are too low throughput and are difficult to integrate into SBS formats for high throughput screening (HTS) workflows. Contrastingly, bulk cell sorting methods such as MACS are easily integrated into HTS formats but still have limited parallelization, high sample consumption, and poor performance with rare cell types. The inherent tradeoff of precision and scalability impedes immunological R&D and clinical translation efforts. Through primary market research interviews using I Corp techniques, researchers have voiced their dissatisfaction with the current cell isolation tools saying they are a ?major roadblock in our work on phenotypic analysis.? We reasoned that a product that can highly parallelized and efficient cell purification in an SBS well plate format would alleviate these bottlenecks and accelerate clinical translation. Leveraging a core technology known as ratcheting cytometry, the objective of this proposal to develop a ?smart plate? which can achieve integrated cell purification in a 96 well plate format. In preliminary studies, ratcheting cytometry has demonstrated quantitative cell sorting capacity and capacity to self-assemble cells into a single cell format. In phase I, ratcheting cytometry will be compared to the gold standard of MACS in isolating CD 14+ monocytes. In parallel, a scaled ratcheting cytometry instrument will be developed to accommodate whole well plate formats. Finally, a prototype ratcheting cytometry smart plate will be developed and tested on the scaled instrument. Phase II will focus on developing a repeatable smart plate design with established QC metrics and instrument integration with robotic fluid handlers.